Evidence for the involvement of the NADPH oxidase enzyme complex in the optimal accumulation of platelet-activating factor in the human cell line PLB-985

Tejindervir S. Hiran, Mary C. Dinauer, Christopher Johnson, Keith L. Clay, Jeffrey B. Travers

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Platelet-activating factor (PAF) is an early product of the inflammatory environment, influencing development and resolution of inflammation. Its production is greater in neutrophils and macrophages, which predominantly synthesize 1-alkyl sn-2 acetyl glycerophosphocholine (GPC) than in non-granulocytes (B cells and endothelial cells), which lack a respiratory burst and synthesize 1-acyl sn-2 acetyl GPC as their major PAF species. This study investigated whether the respiratory burst was responsible for the quantitative and qualitative differences in sn-2 acetyl GPC species generation by neutrophils and macrophages versus those cells lacking the NADPH oxidase complex. The myeloid cell line PLB-985 (capable of differentiation into neutrophils) was used to test this hypothesis, since these cells had previously been generated with a non-functional respiratory burst (X-CGD PLB-985). Differentiated PLB-985 cells underwent a large respiratory burst in response to PMA (phorbol ester), and smaller respiratory bursts in response to A23187 (calcium ionophore), and the bacterial polypeptide fMLP (receptor mediated activation). Concurrently, treated cells were assessed for production of 1-hexadecyl and 1-palmitoyl sn-2 acetyl GPC species by gas chromatography/mass spectrometry. Neither cell type generated these lipid species in response to PMA, but both cell types generated equal levels of sn-2 acetyl GPC in response to A23187, with five times more 1-hexadecyl than 1-palmitoyl species. Upon fMLP activation, X-CGD PLB-985 cells produced significantly less 1-hexadecyl and 1-palmitoyl sn-2 acetyl GPC in comparison to the wild-type PLB-985 cells. These findings suggest phagocytic oxidant production by NADPH oxidase is not essential for sn-2 acetyl GPC generation, but appears important for optimal production of PAF in response to some stimuli.

Original languageEnglish (US)
Pages (from-to)305-315
Number of pages11
JournalProstaglandins and Other Lipid Mediators
Volume66
Issue number4
DOIs
StatePublished - Dec 1 2001

Fingerprint

NADPH Oxidase
Platelet Activating Factor
Respiratory Burst
Cells
Cell Line
Macrophages
Enzymes
Chemical activation
Neutrophils
Calcium Ionophores
Endothelial cells
Calcimycin
Phorbol Esters
Oxidants
Gas chromatography
Mass spectrometry
Lipids
Peptides
Myeloid Cells
Gas Chromatography-Mass Spectrometry

Keywords

  • Chronic granulomatous disease
  • fMLP
  • Granulocytes
  • NADPH oxidase
  • Platelet-activating factor

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Cite this

Evidence for the involvement of the NADPH oxidase enzyme complex in the optimal accumulation of platelet-activating factor in the human cell line PLB-985. / Hiran, Tejindervir S.; Dinauer, Mary C.; Johnson, Christopher; Clay, Keith L.; Travers, Jeffrey B.

In: Prostaglandins and Other Lipid Mediators, Vol. 66, No. 4, 01.12.2001, p. 305-315.

Research output: Contribution to journalArticle

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